LOCUS       Exported                6659 bp ds-DNA     circular SYN 13-DEC-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6659)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jan 28, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6659
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             947..2155
                     /codon_start=1
                     /product="NM_000967 with G111A silent mutation"
                     /label=human RPL3
                     /translation="MSHRKFSAPRHGSLGFLPRKRSSRHRGKVKSFPKDDPSKPVHLTA
                     FLGYKAGMTHIVREVDRPGSKVNKKEVVEAVTIVETPPMVVVGIVGYVETPRGLRTFKT
                     VFAEHISDECKRRFYKNWHKSKKKAFTKYCKKWQDEDGKKQLEKDFSSMKKYCQVIRVI
                     AHTQMRLLPLRQKKAHLMEIQVNGGTVAEKLDWARERLEQQVPVNQVFGQDEMIDVIGV
                     TKGKGYKGVTSRWHTKKLPRKTHRGLRKVACIGAWHPARVAFSVARAGQKGYHHRTEIN
                     KKIYKIGQGYLIKDGKLIKNNASTDYDLSDKSINPLGGFVHYGEVTNDFVMLKGCVVGT
                     KKRVLTLRKSLLVQTKRRALEKIDLKFIDTTSKFGHGRFQTMEEKKAFMGPLKKDRIAK
                     EEGA"
     CDS             2171..2194
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2211..2229)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2255..2479
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2525..2953
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2967..3297
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3148..3283
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3364..4158
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4332..4453
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4502..4518)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4526..4542
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4550..4580)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4595..4616
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4904..5492)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5663..6523)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6524..6628)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcaccatgt ctcacagaaa
      961 gttctccgct cccagacatg ggtccctcgg cttcctgcct cggaagcgca gcagcaggca
     1021 tcgtgggaag gtgaagagct tccctaagga tgacccatcc aagccggtcc acctcacagc
     1081 cttcctggga tacaaggctg gcatgactca catcgtgcgg gaagtcgaca ggccgggatc
     1141 caaggtgaac aagaaggagg tggtggaggc tgtgaccatt gtagagacac cacccatggt
     1201 ggttgtgggc attgtgggct acgtggaaac ccctcgaggc ctccggacct tcaagactgt
     1261 ctttgctgag cacatcagtg atgaatgcaa gaggcgtttc tataagaatt ggcataaatc
     1321 taagaagaag gcctttacca agtactgcaa gaaatggcag gatgaggatg gcaagaagca
     1381 gctggagaag gacttcagca gcatgaagaa gtactgccaa gtcatccgtg tcattgccca
     1441 cacccagatg cgcctgcttc ctctgcgcca gaagaaggcc cacctgatgg agatccaggt
     1501 gaacggaggc actgtggccg agaagctgga ctgggcccgc gagaggcttg agcagcaggt
     1561 acctgtgaac caagtgtttg ggcaggatga gatgatcgac gtcatcgggg tgaccaaggg
     1621 caaaggctac aaaggggtca ccagtcgttg gcacaccaag aagctgcccc gcaagaccca
     1681 ccgaggcctg cgcaaggtgg cctgtattgg ggcatggcat cctgctcgtg tagccttctc
     1741 tgtggcacgc gctgggcaga aaggctacca tcaccgcact gagatcaaca agaagattta
     1801 taagattggc cagggctacc ttatcaagga cggcaagctg atcaagaaca atgcctccac
     1861 tgactatgac ctatctgaca agagcatcaa ccctctgggt ggctttgtcc actatggtga
     1921 agtgaccaat gactttgtca tgctgaaagg ctgtgtggtg ggaaccaaga agcgggtgct
     1981 caccctccgc aagtccttgc tggtgcagac gaagcggcgg gctctggaga agattgacct
     2041 taagttcatt gacaccacct ccaagtttgg ccatggccgc ttccagacca tggaggagaa
     2101 gaaagcattc atgggaccac tgaagaaaga ccgaattgca aaggaagaag gagcttggcg
     2161 gccgctcgag gactacaagg acgacgatga caagctctag agggccctat tctatagtgt
     2221 cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     2281 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2341 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2401 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     2461 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     2521 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2581 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2641 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga
     2701 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2761 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2821 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2881 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2941 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3001 ccccaggcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3061 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3121 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3181 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     3241 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     3301 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     3361 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3421 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3481 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3541 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3601 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3661 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3721 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3781 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3841 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     3901 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3961 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4021 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4081 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4141 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4201 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4261 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4321 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4381 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4441 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     4501 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4561 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4621 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4681 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4741 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4801 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4861 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4921 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4981 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5041 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     5101 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5161 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5221 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5281 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5341 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5401 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5461 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5521 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5581 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5641 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5701 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5761 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5821 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5881 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5941 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6001 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6061 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6121 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6181 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6241 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6301 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6361 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6421 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6481 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6541 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6601 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
